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Factors That Predisposing to Persea Americana Mill. to Infections of Botryosphaeriaceae Species in Central Zone of Chile

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Factors That Predisposing to Persea Americana Mill. to Infections of Botryosphaeriaceae Species in Central Zone of Chile Factors that Predisposing to Persea americana Mill. to Infections of Botryosphaeriaceae Species in Central Zone of Chile Ana Luisa Valencia Díaz 2019 Pontificia Universidad Católica de Chile Facultad de Agronomía e Ingeniería Forestal Factors that Predisposing to Persea americana Mill. to Infections of Botryosphaeriaceae Species in Central Zone of Chile Ana Luisa Valencia Díaz Thesis to obtain the degree of Doctor en Ciencias de la Agricultura Santiago, Chile, April 2019 Thesis presented as part of the requirements for the degree of Doctor en Ciencias de la Agricultura, approved by the Thesis Committee _____________________ Dr. Pilar M. Gil, Advisor __________________ Dr. I. Marlene Rosales, Co-Advisor ___________________ Dr. Johanna Martiz _______________________________ Dr. Jorge Saavedra Santiago, April 2019 To Alonso my husband, my daughters Josefa and Antonella for their unconditional support, that allow me growth to professional level. This work was financial supported by Doctoral Scholarship CONICYT-PCHA 21140282, Project CONICYT-PAI 781413002, and Subsole S.A. The Dirección de Investigación y Postgrados of Facultad de Agronomía e Ingeniería Forestal supported mainly in diffusion activities of the main themes of this thesis. Acknowledgements To Dr. Pilar Gil who motivated me to develop this Doctoral thesis, and to continue with the study of plant diseases from a global perspective. To Dr. Pilar Gil, Dr. Marlene Rosales, Dr. Jorge Saavedra, and Dr. Johanna Martiz, for their support and his great capacity academic and professional, allowing the development of this thesis. Recognize the collaboration of Alonso Retamales for his technical support in the field and laboratory. To Francisco González, Gonzalo Vargas, and staff of Subsole S.A., specially to Andrés Link for his technical support in the field and packing. To producers of Fundo Los Agustinos, San Felipe; FullPal S.A. Nogales; Cuatro Palmas S.A., Quillota; Parcela El Río, Ocoa; Agrícola El Roble, Jaururo; Fundo Quilhuica, María Pinto; Parcela La Muralla, Alicahue; Fundo El Cardal, Melipilla; Parcela 147, Illapel; Parcela 5, La Ligua; La Rosa SOFRUCO, Peumo; Agrícola Doña Adriana, El Carmen de Las Rosas, because they allowed us to examine their orchards, and contribute with samples of branches and fruits for the development of this research in two growing seasons. To staff of our university laboratories: Agua y Riego, Fitopatología Molecular, Patología Frutal y Postcosecha, for their technical assistance and allow used their installations during the development of this research. To Dr. Bernardo Latorre, Dr. Gonzalo Díaz, Dr. Enrique Ferrada, and Dr. Karina Elfar, for giving me part of their time for answer and/or helping me to develop techniques related to my thesis and their friendship. To Daniela Cea, Erika Ramirez, Karen Campos, Pablo Morales, and Claudia Rojas, friends, with whom I shared great moments. Contents index Page Chapter 1: General Introduction.…………………………………………...…......... 1 The cultivation of avocado trees in Chile………………..…........... 2 Export of avocados……………………………………………….…. 2 Effect of pathogens in deterioration of the fruit………………….... 3 Pathologies associated with Botryosphaeriaceae species…….... 5 Effect of Botryosphaeriaceae species in growing avocado……… 6 Effect of Botryosphaeriaceae species in quality postharvest……. 9 Hypothesis and objectives ………………………...………….….............. 11 References………………..………….……………………………………... 12 Chapter 2: Characterization and Pathogenicity of Botryosphaeriaceae Species Obtained from Avocado Trees with Branch Canker and Dieback, and from Avocado Fruit with Stem End Rot in Chile…………..……………………………... 19 Abstract……..……………………………..……………………................ 20 Introduction………………………………….…………………………….. 21 Materials and methods……………………………………………….….. 25 Field sampling………………………..………..…............…………... 25 Fungal isolation…...………………………………….......……….….. 26 Morphological characterization………………………………...…..... 27 Temperature effects on mycelial growth …………………….......... 27 DNA extraction, PCR amplification and sequencing……............... 27 Phylogenetic analyses…………………………………..........……… 29 Pathogenicity testing..………………………………………..........… 29 Results……………………………………………..…………................... 32 Field sampling……………………………………….........…………... 32 Fungal isolation……………..……………...……….........……….….. 32 Morphological characterization…….………………….....………..... 32 Temperature effects on mycelial growth …………………….....….. 35 DNA extraction, PCR amplification and sequencing……...…….... 35 Phylogenetic analyses……..…….…..………..........…………......... 36 Pathogenicity testing..……………………..……..........…………...... 37 Discussion………………….……………..…….........………….............. 39 Acknowledgements……….……………………………………………… 45 Funding…………………………………………………………………….. 45 Literature cited..…………………….…………………………………...... 45 Chapter 3: Effect of Edaphoclimatic Conditions, Planting and Orchard Management in Branch Canker, Dieback and Stem End Rot in Chilean Avocado Orchards............................................................................................................. 69 Abstract…...………………………………………………………............. 70 Introduction……………………………………………………………....... 71 Materials and methods……………………………………………….…... 76 Field sampling………………....………………………………………. 77 Isolation, culture, and identification……………….....………………. 77 Characterization of orchards…………………………………….…… 78 Statistical analysis…………………………….……………………….. 79 Results………………………………….…………………………............. 82 Field sampling………………….………………………………………. 82 Fungal isolation……………….……………………………………….. 82 Analysis of variance……………….…………………………………... 83 Multivariate analysis…………………….…………………………….. 84 Discussion..……………………….……………………………………..... 88 Acknowledgements……..………………………………………………… 93 Literature cited..………….……………………………………………...... 93 Chapter 4: Predictive model to segregate fruit by risk of developing stem end rot in avocados stored for long periods...…………………………………………… 104 4.1. Abstract…………………………………………………………………….. 105 4.2. Introduction……………………………………………………….……...... 106 4.3. Materials and methods……………………………………………………. 110 Field sampling………………....………………………………………. 110 Isolation, culture, and identification……………….....………………. 110 Characterization of orchards…………………………………….…… 111 Statistical analysis…………………………….……………………….. 113 4.4. Results……………………………………………………………………… 116 Field sampling………………….………………………………………. 116 Fungal isolation……………….……………………………………….. 116 Statistical analysis…………………………….………………………… 116 4.5. Discussion and conclusion..…………………………………….……...... 120 4.6. Acknowledgements……………………………………………………….. 124 4.7. References………………………………………………………………… 124 Chapter 5: General Discussion……………………………………………………… 134 References………………………………………………………………… 137 Chapter 6: Conclusions……….……………………………………………………… 139 Chapter 7: Annexes…………………………………………………………………... 140 Annex 7.1. Valencia A. L, and Gil P.M. 2015. Conditions detected in avocado orchards to develop canker dieback caused by Botryosphaeriaceae species in Chile. VIII World Avocado Congress. Lima, Perú. 7.1.1. Abstract.......................................................................................... 140 7.1.2. Poster............................................................................................. 142 7.1.3. Proceeding..................................................................................... 143 Annex 7.2. Valencia A.L., Gil P.M., Rosales M., Saavedra-Torrico J., Martiz J., and Link A. 2016. Estudio de condiciones predisponentes en Persea americana variedad Hass, para el desarrollo de cancrosis, muerte regresiva y pudrición peduncular causadas por Botryosphaeriaceae detectadas en Chile. 67° Congreso Agronómico y 14° Sochifrut. Santiago, Chile. 7.2.1. Abstract.......................................................................................... 148 Annex 7.3. Valencia A.L., Gil P.M., Rosales M., Saavedra-Torrico J., Link A., and Gonzalez F. 2016. Seminario Proyecto CONICYT+PAI 781413002: Estudio de Factores asociados y especies causantes de enfermedad en la madera y pudrición peduncular en huertos de paltos Hass de Chile. Santiago, Chile. 7.3.1. Invitation....................................................................................... 150 7.3.2. Program………………………………………………..……………… 152 Annex 7.4. Valencia A.L., Gil P.M., and Rosales M. 2017. Caracterización y patogenicidad de especies de Botryosphaeriaceae obtenidas desde madera y frutos de Persea americana en huertos chilenos. XXV Congreso SOCHIFIT, XIX Congreso ALF y LVII APS Caribbean división meeting. Chillán, Chile. 7.4.1. Abstract......................................................................................... 153 7.4.2. Prize…………………………………………………………………… 155 Annex 7.5. Avocado Brainstorming. South Africa. 7.5.1. Poster. Valencia A.L., Gil P.M., and Rosales M. 2018. Characterization and Pathogenicity of Botryosphaeriaceae Species obtained from Wood and Fruits of Persea americana in Chilean Orchards. ........................................................................ 156 7.5.2. Poster. Valencia A.L., Gil P.M., Rosales M., and Saavedra-Torrico J. 2018. Factors detected in avocado orchards to develop branch canker, dieback and stem end rot caused by Botryosphaeriaceae species in Chile. ........................................................................... 157 Annex 7.6. Valencia A.L., Gil P.M., Rosales M., Saavedra-Torrico J., Martiz J., and Link A. 2018. Botryosphaeriaceae en palto: agentes causales de enfermedades de la madera y pudrición peduncular del fruto. Redagrícola Chile. Especial paltos
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